1887

Abstract

Summary

The replication of influenza virus RNA was monitored by RNA-RNA hybridization with subsequent analysis of hybrid duplexes, as well as by immunosorbtion of viral nucleocapsids from extracts of [H]uridine-labelled cells followed by the isolation and characterization of nucleocapsid-associated RNA. The nucleocapsid-associated RNA preparations contained mostly negative-strand genomic RNA. Electrophoresis of the hybrid RNA duplexes or single-stranded nucleocapsid-associated RNA in polyacrylamide gel revealed an ‘early’ replication pattern, with a predominance of the NP and NS gene segments, in cells labelled from 0 to 1 h post-infection. At later stages of infection the pattern changed to the ‘late’ one, with the M gene segment in excess of NS, and the NP gene no longer predominant. Cycloheximide added as late as 2 or 3 h post-infection suppressed RNA replication. Moderate concentrations of cycloheximide inhibited the replication of NS and NP gene segments to a lesser degree than the replication of the other RNA segments, thus restoring the ‘early’ replication pattern. Cycloheximide treatment resulted in a slight increase in the percentage of positive strands in nucleocapsid-associated RNA. The role of protein synthesis in the transition from the ‘early’ to the ‘late’ pattern of influenza virus RNA synthesis is discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-68-11-2879
1987-11-01
2022-01-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/68/11/JV0680112879.html?itemId=/content/journal/jgv/10.1099/0022-1317-68-11-2879&mimeType=html&fmt=ahah

References

  1. Barrett T., Wolstenholme A. J., Mahy B. W. J. 1979; Transcription and replication of influenza virus RNA. Virology 98:211–225
    [Google Scholar]
  2. Bautle J. A., Maxwell I. H., Hahn W. E. 1976; Specificity of oligo(dT)-cellulose chromatography in the isolation of polyadenylated RNA. Analytical Biochemistry 72:413–427
    [Google Scholar]
  3. Bean B. Y., Sriram G., Webster R. G. 1979; Electrophoretic analysis of iodine-labeled influenza virus RNA segments. Analytical Biochemistry 102:228–232
    [Google Scholar]
  4. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  5. Enami M., Fukuda R., Ishihama A. 1985; Transcription and replication of eight RNA segments of influenza virus. Virology 142:68–77
    [Google Scholar]
  6. Ghendon Y. Z., Markushin S. G., Klimov A. I., Hay A. J. 1982; Studies of fowl plague virus temperature-sensitive mutants with defects in transcription. Journal of General Virology 63:103–111
    [Google Scholar]
  7. Hay A. J., Lomniczi B., Bellamy A. R., Skehel J. J. 1977; Transcription of the influenza virus genome. Virology 83:337–355
    [Google Scholar]
  8. Ito Y., Joklik W. K. 1972; Temperature-sensitive mutants of reovirus. I. Patterns of gene expression by mutants of group C, D and E. Virology 50:189–201
    [Google Scholar]
  9. Kaverin N. V., Varich N. L., Sklyanskaya E. I., Amvrosieva T. V., Petrik J., Vovk T. C. 1983; Studies on heterotypic interference between influenza A and B viruses: a differential inhibition of the synthesis of viral proteins and RNAs. Journal of General Virology 64:2139–2146
    [Google Scholar]
  10. Kingsbury D. W. 1966; Newcastle disease virus RNA. II. Preferential synthesis of RNA complementary to parental viral RNA by chick embryo cells. Journal of Molecular Biology 18:203–214
    [Google Scholar]
  11. Kolakofsky D., Boy De La Tour E., Delius H. 1974; Molecular weight determination of Sendai and Newcastle disease virus RNA. Journal of Virology 13:261–268
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  13. Mahy B. W. J. 1983; Mutants of influenza virus. In Genetics of Influenza Viruses pp. 192–254 Palese P., Kingsbury D. W. Edited by Wien & New York: Springer-Verlag;
    [Google Scholar]
  14. Mark G. E., Taylor J. M., Broni B., Krug R. M. 1979; Nuclear accumulation of influenza virus RNA transcripts and the effect of cycloheximide, actinomycin D and a-amanitin. Journal of Virology 29:744–752
    [Google Scholar]
  15. Meier-Ewert H., Compans R. W. 1974; Time course of synthesis and assembly of influenza virus proteins. Journal of Virology 14:1083–1091
    [Google Scholar]
  16. Pons M. 1975; Influenza virus RNA(s). In Influenza Viruses and Influenza pp. 145–170 Kilbourne E. D. Edited by New York, San Francisco & London: Academic Press;
    [Google Scholar]
  17. Scholtissek C., Rott R. 1970; Synthesis in vivo of influenza virus plus and minus strand RNA and its preferential inhibition by antibiotics. Virology 40:989–996
    [Google Scholar]
  18. Shapiro G. I., Gurney T., Krug R. M. 1987; Influenza virus gene expression: control mechanisms at early and late times of infection and nuclear-cytoplasmic transport of virus-specific RNAs. Journal of Virology 61:764–773
    [Google Scholar]
  19. Skehel J. J. 1973; Early polypeptides synthesis in influenza virus-infected cells. Virology 56:394–399
    [Google Scholar]
  20. Sklyanskaya E. I., Varich N. L., Amvrosieva T. V., Kaverin N. V. 1985; Virions and intracellular nucleocapsids produced during mixed heterotypic influenza infection of MDCK cells. Journal of Virology 53:679–683
    [Google Scholar]
  21. Smith F. G. L., Hay A. J. 1982; Replication of the influenza virus genome. Virology 118:96–108
    [Google Scholar]
  22. Spear P. G., Roizman B. 1972; Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpes virion. Journal of Virology 9:143–159
    [Google Scholar]
  23. Varich N. L., Lukashevich I. S., Kaverin N. V. 1979; Newcastle disease virus-specific RNA: an analysis of 24S and 35S RNA transcripts. Acta virologica 23:273–283
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-68-11-2879
Loading
/content/journal/jgv/10.1099/0022-1317-68-11-2879
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error